Transcription and cap trimethylation of a nematode spliced leader RNA in a cell-free system.
Maturation of a fraction of mRNAs in nematodes involves the acquisition of a common 5' terminal spliced leader sequence derived from a nonpolyadenylylated spliced leader RNA by trans splicing. We have developed a cell-free system prepared from Ascaris lumbricoides embryos that accurately and efficiently synthesized the spliced leader RNA of A. lumbricoides. Transcription of the spliced leader RNA was catalyzed by RNA polymerase II, and the majority of the spliced leader RNAs synthesized in vitro possessed a trimethylguanosine cap structure identical to that found on in vivo-synthesized spliced leader RNA.
Maroney
P A
PA
Department of Molecular Biology and Microbiology, Case Western Reserve University, Cleveland, OH 44106.
Hannon
G J
GJ
Nilsen
T W
TW
D000818:Animals; D001200:Ascaris / Q000196:embryology / Q000235:genetics*; D001483:Base Sequence; D002474:Cell-Free System; D004625:Embryo, Nonmammalian / Q000378:metabolism; D008745:Methylation; D008969:Molecular Sequence Data; D009710:Nucleotide Mapping; D012315:RNA Caps / Q000235:genetics* / Q000378:metabolism; D012319:RNA Polymerase II / Q000378:metabolism; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics*; D012342:RNA, Small Nuclear / Q000235:genetics; D014158:Transcription, Genetic
D012315:RNA Caps; D012333:RNA, Messenger; D012342:RNA, Small Nuclear; D012319:RNA Polymerase II